全 文 ：第 25 卷第 4期
2008 年 7月
中 国科学院 研究生院 学报
Journal of the Graduate School of the Chinese Academy of Sciences
Vol.25
July
　No.4
2008
Article ID:1002-1175(2008)04-0569-07
EXCERPTS OF DISSERTATION
Studies on the chemical constituents and
bioactivities of five Schisandra medicinal
species and Elsholtzia bodinieri
＊
LI Rong-Tao　SUN Han-Dong
(State Key Laboratory of Phytochemistry and Plant Resources in West China , Kunming Institute of Botany ,
Chinese Academy of Sciences , Kunming 650204 , China)
(Received 28 November 2007)
Li RT, Sun HD.Studies on the chemical constituents and bioactivities of five Schisandra medicinal species and Elsholtzia
bodinieri.Journal of the Graduate School of the Chinese Academy of Sciences , 2008 , 25(4):569 ～ 575
Abstract　The chemical constituents and bioactivities of six medicinal plants have been studied.In the
study one hundred and one compounds , including twenty-six new compounds were isolated.Among these
compounds , sixteen unique highly oxidized nortriterpenoids , belonging to an unprecedented new
nortriterpenoid skeleton with a biosynthetically modified eight-membered ring , and unusual
nortriterpenoids or bisnortriterpenoid skeletons , have been isolated from three Schisandra species.
Interestingly , to date , no naturally occurring triterpenoids have been found to have such a highly modified
oxidized norcycloartane skeleton.
The bioactivities of compounds isolated from the genus Schisandra have been tested , including anti-
HIV-1 , anti-inflammatory , and cytotoxities against C8166 , MT-4 and K562 cells activities.SM-10 and
SM-26 were found to show promising anti-HIV-1 activity and the selectivity index were 174.08 and >
25.94 , respectively.
Key words 　Schisandra species , Elsholtzia bodinieri , chemical constituents , anti-HIV activity ,
biogenetic origin
CLC　Q946-33
＊supported by the National Natural Science Foundat ion of China(20402016)
Triterpenoids are the most ubiquitous , nonsteroidal secondary metabolites in terrestrial and marine flora and
fauna.Their presence , even in nonphotosynthetic bacteria , has created interest from both an evolutionary and
functional point of view.Although medicinal uses of the compounds from this class are rather limited , considerable
recent work in this regard strongly indicates their great potential as drugs
[ 1] .Moreover , despite the great diversity
that already exists among the skeletons of triterpenoids , new variants continue to emerge.Some of the new
triterpenoid structures possess novel skeletons and represent unique biosynthetic end products.The majority of
triterpenoids possess the conventional skeleton arising from the cyclization of squalene-2 , 3-epoxide to yield fused
polycyclic products.More unusual are the incompletely cyclized compounds , or those exhibiting cyclization within
the chain , or two consecutive cyclizations rather than the cyclization beginning at one end.While triterpenoids with
rearranged carbocyclic skeletons have been isolated quite frequently , there are some skeletons formed through
extensive oxidation accompanied by various bond cleavages
[ 2 , 3] .
With the aim of identifying new natural compounds with interesting biological activities and of investigating the
occurrence of natural terpenoids that could be used as natural sources of intermediates for the synthesis of high-added
value compounds , we examined the plants of the genus Schisandra grown in Yunnan Province , which belongs to the
economically and medicinally important family Schisandra ceae.
Sixteen highly oxygenated nortriterpenoids(Fig.1), with three unprecedented carbon skeletons , of which we
proposed the names “ schisanartane” , “ schiartane” , and “ 18-norschiartane (Fig.3), have been isolated and
characterized , some of which have potent anti-HIV-1 activity with low toxicity.The schisanartane skeleton
triterpenoids are the main constituents of Schisandra species and are also a unique family of cycloartane derived
triterpenoids because of their octacyclic backbone , which includes a 7 8 5 consecutive carbocycle and more than 12
chiral centers.These unusual ring assembly and highly oxygenated structure features are distinctive from any other
naturally occurring triterpenoids.As a consequence , these structurally complex molecules have brought great interest
and challenges to the chemists for total synthesis and biogenetic studies
[ 4 , 5] .
1 　Investigation on chemical constituents of five Schisandra species and
Elsholtiza bodinieri
　　The chemical constituents and bioactivities of five medicinal Schisandra plants , Schisandra micrantha A.C.
Smith , S .lancifolia(Rehd.et Wils.)A.C.Smith , S.henryi var.yunnanensis var.yunnanensisA.C.Smitt ,
S.Plena A.C.Smith , and S .sphenanthera Rehd.et Wils , have been studied.Eighty seven compounds ,
including twenty-one new compounds were isolated.Their structures have been elucidated on the basis of extensive
spectroscopic studies in conjunction with single-crystal X-ray diffraction analysis.
1.1　Study on chemical constituents from S.micrantha
Our investigation on the chemical constituents of the leaves and stems of S.micrantha resulted in the isolation
of ten novel compounds , together with twenty-five known ones.Five novel nortriterpenoids , micrandilactones A ,
D-G(1 ～ 5) are the first examples of structurally unique highly oxidized norcycloartane skeleton with a
biosynthetically modified eight-membered ring D isolated from the family Schisandraceae.Two novel
nortriterpenoids, micrandilactones B-C(6 , 7), represent another unprecedented highly oxidized norcycloartane
skeleton(Fig.1).Micranoic acids A and B(8 ,9)belong to a new group of octanortriterpenoids in which the entire
C-17 side chains have been lost(Fig.2).Micrantherin A is a new dibenzocyclooctadiene lignan[ 6 ～ 10] .
1.2　Investigation on chemical constituents of S.lancifolia
In our present study , fifteen compounds , including five novel ones , were isolated from the leaves and stems of
S .lancifolia .The skeletal type displayed by lancifodilactone A(10)is noticeable for it represents a new group of
highly oxygenated bisnortriterpenoid.Lancifodilactones B-E(11 ～ 14)is a novel compound possessing the same
schisanartane skeleton with that of micrandilactone A , of which was firstly isolated from S .micrantha
(Fig.1)[ 11 ,12] .
1.3　Phytochemical study on S.henryi var.yunnanensis
The investigation of the leaves and stems of S .henryi var.yunnanensis led to the isolation of sixteen
compounds.Among them , four novel nortriterpenoids , henridilactones A-D(15 ～ 18), are members of a unique
unusual highly oxidized schisanartane skeletal class structurally related to micrandilactone A(Fig.1)[ 13] .
570 中国科学院研究生院学报 第 25 卷
Fig.1　Novel nortriterpenoids with new skeletons
　
Fig.2　Unusual Octanortriterpenoids and 18 , 19-seco-ursane glycosides
　
Fig.3　Three new skeletons
　
1.4　Research on Chemical Constituents from S.plena and S.sphenanthera
The chemical constituents of the leaves and stems of S .plena are described for the first time.This
investigation has resulted in the isolation of a new sesquiterpenoid , plenoxide.In addition , thirteen known
compounds , including sesquiterpenoids , coumarins , flavanones , triterpenoids and steroids were also isolated.The
detailed analysis of 2D NMR data also led us to conclude that the chemical shifts of earlier compounds similar to
bullatantriol need revision
[ 14] .
A new phenolic glycoside , 4 ,6-dihydroxyphenyl-1-butanone-2-β-O-glucopyranoside , along with seven known
compounds were isolated from the leaves and stems of S.sphenanthera.
1.5　Investigation of chemical constituents from Elsholtzia bodinieri
We have investigated the aerial parts of Elsholtzia bodinieri and herein report the isolation of fifteen
compounds , including eleven flavanones and four triterpenoids.Among them , there are two novel 18 , 19-seco-
ursen-28-oic acid-3-O-β-D-glucopyranoside triterpenoids , bodiniosides A(19) and B(20)(Fig.2)[ 15] .In
addition , three new flavanones are determined as eriodictyol 7-O-(6″-feruloyl)-β-D-glucopyranoside , eriodictyol 7-
O-(6″-isoferuloyl)-β-D-glucopyranoside , and luteolin 7-O-(6″-isoferuloyl)-β-D-glucopyranoside.
571　第 4 期 LI Rong-Tao , SUN Han-Dong:Studies on the chemical constituents and bioactivities of five Schisandra medicinal… 　　 　
1.6　Structure elucidation of micandilactone A
Table 1　1H and 13 CNMR assignments and HMBC correlations of 1a
No. δH(mult , J , Hz) δC HMBC(1H-13C)
1 4.22(d , 6.3) 81.4 3, 10 , 19
2α 2.74(d , 18.6) 35.0 1, 3 , 10
2 β 2.93(dd , 6.3 , 18.6) 3
3 175.2
4 83.9
5 2.47(dd , 4.2 , 13.4) 58.3 4, 10
6α 2.09(m) 36.4 7, 8
6 β 2.21(overlap) 5, 7
7 4.51(dd , 9.3 , 10.1) 67.8 5, 16
8 2.99(d , 10.1) 59.7 6, 7 , 9 , 15, 16 , 19
9 82.2
10 95.6
11α 1.79(m) 42.3 13
11β 1.98(m) 9, 19
12α 1.67(m) 32.6 13 , 17
12β 1.98(overlap) 14
13 49.3
14 3.31(s) 54.1 13 , 15 , 16 , 17 , 18 , 20 ,
22
15 99.7
16 207.4
17 220.7
18 1.58(s) 30.8 12 , 13 , 14, 17
19α 2.52(ABd, 15.8) 41.8 9
19β 2.23(ABd, 15.8) 9, 10 , 11
20 80.2
21 1.77(s) 18.9 17 , 20 , 22
22 75.5
23 4.99(d , 1.5) 76.8 14 , 20 , 22, 24
24 5.42(dd , 1.5 , 2.0) 75.2 23 , 26
25 3.26(m) 42.5 26
26 177.5
27 1.17(d , 7.1) 7.8 24 , 25 , 26
29 1.24(s) 27.7 4, 5 , 30
30 1.04(s) 20.8 4, 5 , 29
20-OH 5.90(s) 17 , 20 , 21, 22
22-OH 7.56(s) 20
　　a Data were recorded in C5D5N on Bruker AM-400 MHz spectrometer(1H , 13C)and Bruker
DRX-500MHz spectrometer (HMBC), chemical shifts(δ)were in 10-6.
　　Micrandilactone A(1)(Fig.4)
crystallized as colorless prisms , has
the molecular formula of C29H36O12
as deduced by its HREI MS (found
576.2178 , calcd 576.2207 ),
requiring 12 degrees of unsaturation.
The IR spectrum showed absorptions
at 3439cm
-1
and 1776cm
-1 ,
revealing the presence of hydroxyl
and γ-lactone groups[ 16] .The 1H
NMR(Table 1)spectrum exhibited
signals due to four tertiary methyls
and a secondary methyl.The 13 C
NMR spectrum indicated that 1
contained two ester groups , two
carbonyl groups , seven quaternary
carbons including six oxygenated
ones , eight methines including four
oxygenated ones , five methylenes
and five methyls , which suggested a
highly oxygenated triterpene
skeleton.
Fig.4　Structure of
micandilactone A
　　Interpretation of HMBC data showed the following correlations(Table 1):Me-29(δ1.24 , s)and Me-30
(δ1.04 , s)with C-4 and C-5;H-1(δ4.22), H2-2(δ2.74 2.93)and H-5(δ2.47)with C-10;H-1 and H2-
2 with an ester group at C-3;H-8(δ2.99)with C-9 and C-19;H2-19(δ2.23 2.52)with C-9 and C-10.This ,
along with two proton spin-systems deduced from COSY correlations , H-1-H-2 and H-5-H-8 , led to the
establishment of partial structure 1a(Fig.5).
A methyl singlet resonance at δ1.58 corresponding to Me-18 showed HMBC cross peaks with a quaternary
carbon(C-13)and with C-12 , C-14 and C-17 , which required that C-12 , C-14 and C-17 all be attached to the
carbon(C-13)bearing the methyl group.This was confirmed by the observations of correlations between the methine
at δ3.31(H-14 , s)and C-13 , C-17 and C-18.Another methyl singlet resonance at δ1.77(Me-21)also showed
HMBC correlations with the other two oxygenated quaternary carbons(C-20 and C-22)and a ketone carbon(C-17)
suggested that the quaternary carbon (C-20)bearing the methyl group (Me-21)was situated between C-17 and
572 中国科学院研究生院学报 第 25 卷
C-22.Furthermore , correlations of H-14 with C-20 and C-22 established the connection of C-14 with C-22.
Additional COSY correlations(H2-11-H2-12)not only established the attachment of C-11-C-12 but also gave rise to
partial structure 1b.The third fragment 1c , was assigned by a continuous sequence from C-23 to C-26 and Me-27
deducing from COSY and HMBC spectra , as well as by the characteristic IR spectralγ-lactone group absorption
(1776 cm-1)[ 16] .A hydroxyl group (δ5.90)was assigned as 20-OH for its cross peaks with C-17 , C-20 , C-22
andMe-21.In the same manner , another hydroxyl group (δ7.56)was proven to be located at C-22 by its
correlation to C-20.Moreover , HMBC correlations observed between H2-19 and C-9 , C-11 and between H2-11(δ
1.79 1.98)and C-9 , C-19 allowed the combination of 1a and 1b to afford 1d.Further more , correlations of H-23
to C-14 , C-20 and C-22 required direct connection of C-23 with C-22 and permitted fragments 1c and 1d to be
joined to one another to get 1e(Fig.5).
Fig.5　Structural fragments of 1
　
Up to now , the above NMR spectroscopic data analysis has elucidated the constitution of partial structure 1e.
However , no additional HMBC connectivities necessary for constructing the structure of 1 were observed among the
three oxygenated methines(C-1 , C-7 and C-24).In addition , as C-3 , C-4 , C-9 , C-10 , C-15 and C-16 were
quaternary carbons , which was not possible to determine the correct connections among these carbons.
Since it was difficult to elucidate the complete structure of 1 only by NMR spectroscopic analysis , the crystals
were submitted to a single crystal X-ray diffraction.A view of the solid-state conformation was provided in Figure 6.
It indicated that 1 had suffered an oxidative cleavage between C-3 and C-4 , followed by lactonization to give rise to
a five-membered lactone ring A and a tetrahydrofuran ring B.C-9 connected to C-15 through an oxygen bridge with
the hydroxyl group located at C-7.Signal due to the angular methyl attached to C-14(Me-28)was obviously absent
in the case of 1 , which suggested that Me-28 had suffered an oxidation to form a carboxylic group , followed by the
loss of CO2.Thus the basic skeleton of 1 was elucidated as 3 ,4:9 ,10-seco-14-norcycloartane.
The relative stereochemistry of 1 was determined by a 2D ROESY experiment and was confirmed unequivocally
by X-ray crystallographic data.Stereochemically , Me-29 was biogenetically α-oriented , Me-30 and Me-21 were β-
oriented.ROESY correlations for Me-30 H-1 , Me-29 H-5 , H-5 H-7 , 20-OH H-24 and H-24 H-23 , H-25
indicated that H-1 was β-orientation , H-5 , H-7 , H-23 , H-24 and H-25 were the same α-orientations.Me-18 and
H-14 showed mutual correlations , but no cross peaks with H-23 , suggested that H-14 and Me-18 were β-
orientations.H-8 was suggested to be β-oriented considering the coupling constant(1H , d , J 7 ,8 =10.1Hz).The
stereochemistry of the four quaternary carbons , C-9 , C-10 , C-15 and C-22 was deduced as R , S , R and S
configuration , respectively , by X-ray diffraction study.
573　第 4 期 LI Rong-Tao , SUN Han-Dong:Studies on the chemical constituents and bioactivities of five Schisandra medicinal… 　　 　
Fig.6　X-ray structure of 1 showing relative configuration
2　Bioactive investigation
The anti-HIV activities and cytotoxicities of compounds isolated from the genus Schisandra were tested by
microtiter syncytium formation infectivity assay , including cytotoxicity in C8166 and MT-4 cells , inhibition of
syncytium formation in HIV-1ⅢB infected C8166 cells , and protective effect in HIV-1ⅢB infected MT-4 host cells.
Among these compounds , SM-10 possessed cytotoxicity with CC50 value of 22.63 μg mL on tested human T cell
leukemia cell line C8166 at the assayed doses , demonstrated potent anti-HIV-1 activity with EC50 value of 0.13
μg mL(selectivity index:174.08), and exerted protective activity on HIV-1ⅢB infected MT-4 host cells with a
selectivity index of 43.27.SM-26 possessed minimal cytotoxicity(CC50>200μg mL), and the inhibitory activity on
HIV-1ⅢB induced syncytium formation was EC50=7.71μg mL.The selectivity index was >25.94.In addition , SM-
26 exerted its potent protective activity on HIV-1ⅢB infected MT-4 host cells with a selectivity index of >425.5 at
the concentration of 0.47μg mL.
This result is encouraging and warrants further structural modification to both decrease cytotoxicity and increase
antiviral inhibitory activities.The potent anti-HIV-1 activity and unique structural features of SM-10 make it as
promising lead compound for the preparation of anti-HIV drug.Further anti-HIV evaluation is in progress to better
define the anti-HIV potencies of SM-10.Such a study would provide valuable information on the therapeutic
development of new generation for anti-HIV drug.
3　Possible biogenetic origin of schisanartane , schiartane and 18-nor-schi-
artane
　　Since sixteen novel highly oxygenated compounds with three unique skeletons isolated from three Schisandra
species have not previously been encountered in nature , we proposed the name schisanartane , schiartane and 18-
nor-schiartane for the three different types of skeletons.The concurrence of these compounds among three species
prompted us to ponder rationally the inherent correlation among three new skeletons and the known cycloartane.
Orienting these novel compounds in a triterpenic perspective reveals the natural context among schisanartane ,
schiartane , 18-nor-schiartane and the known cycloartane skeletons.
Here we proposed four plausible hypotheses for the biogenesis of three new skeletons:(a)possible biogenesis
of schiartane from cycloartane , (b)biogenetic origin of the new 18-nor-schiartane type , (c)plausible biogenesis of
schisanartane , (d)the interrelationship and a further stereochemical inference among the schisanartanes[ 8] .
574 中国科学院研究生院学报 第 25 卷
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五种五味子属药用植物及东紫苏的化学成分和生物活性
李蓉涛　孙汉董
(中国科学院昆明植物研究所 ,昆明 650204)
摘　要 　对 5 种五味子属(Schisandra)药用植物(小花五味子 Schisandra micrantha , 狭叶五味子 S.
lancifolia ,滇翼梗五味子 S .henryi var.yunnanensis var.yunnanensis ,复瓣黄龙藤 S .Plena ,华中五味子
S .sphenanthera)和唇形科植物东紫苏(Elsholtzia bodinieri)的化学成分进行了研究 ,从中共分离鉴定了
101个化合物 ,26个为新化合物.首次从 3种五味子属植物中发现了 3种高度氧化的新奇骨架类型.此
外 ,还发现了 18 , 19-seco-乌索酸型苷和 17 ,20断裂并失去 17位侧链的羊毛甾烷型和环阿尔廷型八降三
萜等新的骨架类型.
对分离得到的部分化合物进行了体外抗 HIV-1 、急毒 、抗炎和抗肿瘤活性实验 ,发现部分化合物具
有抗HIV-1活性.其中化合物 SM-10和SM-26具有显著的抑制 HIV-1病毒活性 ,选择指数分别为 174.08
和>25.04 ,且毒性较小 ,作为作用于病毒与细胞结合和融合靶点的小分子化合物具有重要的意义.
关键词　五味子属 ,东紫苏 ,化学成分 ,抗HIV活性 ,生源途径
李蓉涛:博士研究生在读期间 ,发表多篇论文 ,荣获 2005年度中国科学院优秀博士学位论文及 2006年度全国优秀百篇博士学位论文奖.
导师孙汉董研究员:主要从事药用和天然香料的植物资源 、植物化学以及天然药物研发的基础和应用基础研究.
575　第 4 期 LI Rong-Tao , SUN Han-Dong:Studies on the chemical constituents and bioactivities of five Schisandra medicinal…